Team:Westminster/Results
Project Results
RESULTS11/09/15
Nanodrop results from plasmid prep
OmcA (1) - 260/280= 1.87 260/230=2.10
TOTAL= 114.5 ng/µl
OmcA (2) - 260/280= 1.88 260/230=2.13
TOTAL= 77.9 ng/µl
CymA (1) - 260/280= 1.86 260/230=2.13
TOTAL= 183.5ng/µl
CymA (2) - 260/280= 1.88 260/230=2.06
TOTAL= 107.1ng/µl
MtrC (1) - 260/280= 1.88 260/230=2.15
TOTAL= 166.7ng/µl
MtrC (2) - 260/280= 1.89 260/230=2.18
TOTAL= 91.1 ng/µl
17/09/15
2 Gels of PCR products extracted yesterday morning (16/09/15) (tube 1, 2, 3, 4, 5, 6, 7, 8, MtrCAB). Gel was run for 40 mins at 100V.
GEL 1
Lane 1 |
Lane 2 |
Lane 3 |
Lane 4 |
Lane 5 |
Lane 6 |
Lane 7 |
Lane 8 |
DNA ladder (10 µl) |
MtrCAB (2 µl + 8 µl) |
Tube 1 (2 µl + 8 µl) |
Tube 2 (2 µl + 8 µl) |
Tube 3 (2 µl + 8 µl) |
Tube 4 (2 µl + 8 µl) |
DNA ladder (10 µl) |
Empty |
Both ladders and MtrCAB were the only lanes with results.
image here
GEL 2
Lane 1 |
Lane 2 |
Lane 3 |
Lane 4 |
Lane 5 |
Lane 6 |
Lane 7 |
Lane 8 |
DNA ladder (10 µl) |
Tube 5 (2 µl + 8 µl) |
Tube 6 (2 µl + 8 µl) |
Tube 7 (2 µl + 8 µl) |
Tube 8 (2 µl + 8 µl) |
DNA ladder (10 µl) |
Empty |
Empty |
Lane 3 was the only lane without results.
image here
Plasmid prep
9 LB bottles were inoculated with amplicillin (100µg/ml) or chlorophenicol (34.5µg/ml) and plasmid pSB1C3 or pSB1A3 from yesterday’s transformation and left to incubate overnight (pSB1C3 =tube 2, 4, 5, 7 and MtrCAB; pSB1A3= tube 1, 3, 6, 8).
Steps |
Tube 1,3,6,8 (Amp) |
Tube 2,5,7 (all Chlr) MtrCAB, |
Tube 4 Chlr |
1. Harvest |
Yes 15mins + extra 5 mins |
Yes 15mins + extra 5 mins |
Yes 15mins + extra 5 mins |
2. Resuspend |
yes |
yes |
yes |
3. Lyse |
yes |
yes |
yes |
4. Precipitate |
yes |
yes |
yes |
5. Bind |
yes |
yes |
yes |
6. Wash (optional) |
yes |
yes |
yes |
7. Wash and Ethanol Removal |
yes |
yes |
yes |
8. Elute |
yes |
yes |
yes |
9. Recover |
yes |
yes |
yes |
Tube 3 (Amp) - resuspended with 340 µl rather than 250 µl
Tube 7 (Chlr) - 360 µl resuspension buffer added rather than 250 µl
Nanodrop results of plasmid prep
Amp 1- 260/280= 1.98 260/230=2.12
TOTAL= 83.0ng/µl
Chlr 2- 260/280= 1.89 260/230=2.11
TOTAL= 151.7ng/µl
Amp 3- 260/280= 1.91 260/230=2.12
TOTAL= 173.5ng/µl
Chlr 4- 260/280= 1.89 260/230=2.18
TOTAL= 92.1ng/µl
Chlr 5- 260/280= 1.93 260/230=2.23
TOTAL= 87.8ng/µl
Amp 6- 260/280= 1.92 260/230=2.15
TOTAL= 129.5ng/µl
Chlr 7- 260/280= 1.91 260/230=2.21
TOTAL= 80.4ng/µl
Chlr 8- 260/280= 1.94 260/230=2.11
TOTAL= 128.4ng/µl
MtrCAB- 260/280= 1.96 260/230=2.28
TOTAL= 38.3ng/µl
Below is the plate that the plasmid DNA that will be freeze dried
|
1 |
2 |
3 |
4 |
A |
1 |
9 |
17 |
25 |
B |
2 |
10 |
18 |
26 |
C |
3 |
11 |
19 |
27 |
D |
4 |
12 |
20 |
|
E |
5 |
13 |
21 |
|
F |
6 |
14 |
22 |
|
G |
7 |
15 |
23 |
|
H |
8 |
16 |
24 |
|
|
1. MtrA pSB1C3 2. MtrB pSB1C3 3. MtrC pSB1C3 4. MtrC (1) 5. MtrC (2) 6. CymA (1) 7. CymA (2) |
8. OmcA 9. OmcA (1) 10. OmcA (2) 11. MtrA-His 12. MtrB-His 13. MtrC-His 14. CymA-His 15. OmcA-His 16- OmcA-CymA(1) |
17. OmcA-CymA (2) 18. MtrCAB 19. Chlr 2 20. Chlr 4 21. Chlr 5 22. Chlr 7 |